Investigation of the pathways related to intrinsic miltefosine tolerance in Leishmania (Viannia) braziliensis clinical isolates reveals differences in drug uptake
In Brazil, cutaneous leishmaniasis is caused predominantly by L. (V.) braziliensis. The few therapeutic drugs available exhibit several limitations, mainly related to drug toxicity and reduced efficacy in some regions. Miltefosine (MF), the only oral drug available for leishmaniasis treatment, is no...
| Main Authors: | , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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Elsevier
2019-12-01
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| Series: | International Journal for Parasitology: Drugs and Drug Resistance |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2211320718301684 |
| _version_ | 1819091287730552832 |
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| author | Caroline R. Espada Rubens M. Magalhães Mario C. Cruz Paulo R. Machado Albert Schriefer Edgar M. Carvalho Valentín Hornillos João M. Alves Angela K. Cruz Adriano C. Coelho Silvia R.B. Uliana |
| author_facet | Caroline R. Espada Rubens M. Magalhães Mario C. Cruz Paulo R. Machado Albert Schriefer Edgar M. Carvalho Valentín Hornillos João M. Alves Angela K. Cruz Adriano C. Coelho Silvia R.B. Uliana |
| author_sort | Caroline R. Espada |
| collection | DOAJ |
| description | In Brazil, cutaneous leishmaniasis is caused predominantly by L. (V.) braziliensis. The few therapeutic drugs available exhibit several limitations, mainly related to drug toxicity and reduced efficacy in some regions. Miltefosine (MF), the only oral drug available for leishmaniasis treatment, is not widely available and has not yet been approved for human use in Brazil. Our group previously reported the existence of differential susceptibility among L. (V.) braziliensis clinical isolates. In this work, we further characterized three of these isolates of L. (V.) braziliensis chosen because they exhibited the lowest and the highest MF half maximal inhibitory concentrations and were therefore considered less tolerant or more tolerant, respectively. Uptake of MF, and also of phosphocholine, were found to be significantly different in more tolerant parasites compared to the less sensitive isolate, which raised the hypothesis of differences in the MF transport complex Miltefosine Transporter (MT)-Ros3. Although some polymorphisms in those genes were found, they did not correlate with the drug susceptibility phenotype. Drug efflux and compartmentalization were similar in the isolates tested, and amphotericin B susceptibility was retained in MF tolerant parasites, suggesting that increased fitness was also not the basis of observed differences. Transcriptomic analysis revealed that Ros3 mRNA levels were upregulated in the sensitive strain compared to the tolerant ones. Increased mRNA abundance in more tolerant isolates was validated by quantitative PCR. Our results suggest that differential gene expression of the MT transporter complex is the basis of the differential susceptibility in these unselected, naturally occurring parasites. Keywords: Leishmania braziliensis, Isolates, Miltefosine, Susceptibility, Uptake, RNAseq |
| first_indexed | 2024-12-21T22:37:20Z |
| format | Article |
| id | doaj.art-e6fdcfd6e9714f0bbed7ebbb1c8e51f4 |
| institution | Directory Open Access Journal |
| issn | 2211-3207 |
| language | English |
| last_indexed | 2024-12-21T22:37:20Z |
| publishDate | 2019-12-01 |
| publisher | Elsevier |
| record_format | Article |
| series | International Journal for Parasitology: Drugs and Drug Resistance |
| spelling | doaj.art-e6fdcfd6e9714f0bbed7ebbb1c8e51f42022-12-21T18:47:56ZengElsevierInternational Journal for Parasitology: Drugs and Drug Resistance2211-32072019-12-0111139147Investigation of the pathways related to intrinsic miltefosine tolerance in Leishmania (Viannia) braziliensis clinical isolates reveals differences in drug uptakeCaroline R. Espada0Rubens M. Magalhães1Mario C. Cruz2Paulo R. Machado3Albert Schriefer4Edgar M. Carvalho5Valentín Hornillos6João M. Alves7Angela K. Cruz8Adriano C. Coelho9Silvia R.B. Uliana10Departamento de Parasitologia, Instituto de Ciências Biomédicas, Universidade de São Paulo, São Paulo, BrazilDepartamento de Biologia Celular e Molecular e Bioagentes Patogênicos, Faculdade de Medicina de Ribeirão Preto, Universidade de São Paulo, Ribeirão Preto, BrazilCentro de Facilidades para Apoio a Pesquisa, CEFAP-USP, Universidade de São Paulo, São Paulo, BrazilServiço de Imunologia, HUPES, Universidade Federal da Bahia, Salvador, BrazilInstituto de Ciências da Saúde, Universidade Federal da Bahia, Salvador, BrazilServiço de Imunologia, HUPES, Universidade Federal da Bahia, Salvador, Brazil; Centro de Pesquisas Gonçalo Moniz, Fiocruz-Bahia, Salvador, BrazilDepartamento de Química Orgánica, Universidad de Sevilla and Centro de Innovación en Química Avanzada (ORFEO-CINQA), Sevilla, SpainDepartamento de Parasitologia, Instituto de Ciências Biomédicas, Universidade de São Paulo, São Paulo, BrazilDepartamento de Biologia Celular e Molecular e Bioagentes Patogênicos, Faculdade de Medicina de Ribeirão Preto, Universidade de São Paulo, Ribeirão Preto, BrazilDepartamento de Parasitologia, Instituto de Ciências Biomédicas, Universidade de São Paulo, São Paulo, BrazilDepartamento de Parasitologia, Instituto de Ciências Biomédicas, Universidade de São Paulo, São Paulo, Brazil; Corresponding author. Departamento de Parasitologia, Universidade de São Paulo, Av. Prof. Lineu Prestes, 1374, CEP 05508-000, São Paulo, Brazil.In Brazil, cutaneous leishmaniasis is caused predominantly by L. (V.) braziliensis. The few therapeutic drugs available exhibit several limitations, mainly related to drug toxicity and reduced efficacy in some regions. Miltefosine (MF), the only oral drug available for leishmaniasis treatment, is not widely available and has not yet been approved for human use in Brazil. Our group previously reported the existence of differential susceptibility among L. (V.) braziliensis clinical isolates. In this work, we further characterized three of these isolates of L. (V.) braziliensis chosen because they exhibited the lowest and the highest MF half maximal inhibitory concentrations and were therefore considered less tolerant or more tolerant, respectively. Uptake of MF, and also of phosphocholine, were found to be significantly different in more tolerant parasites compared to the less sensitive isolate, which raised the hypothesis of differences in the MF transport complex Miltefosine Transporter (MT)-Ros3. Although some polymorphisms in those genes were found, they did not correlate with the drug susceptibility phenotype. Drug efflux and compartmentalization were similar in the isolates tested, and amphotericin B susceptibility was retained in MF tolerant parasites, suggesting that increased fitness was also not the basis of observed differences. Transcriptomic analysis revealed that Ros3 mRNA levels were upregulated in the sensitive strain compared to the tolerant ones. Increased mRNA abundance in more tolerant isolates was validated by quantitative PCR. Our results suggest that differential gene expression of the MT transporter complex is the basis of the differential susceptibility in these unselected, naturally occurring parasites. Keywords: Leishmania braziliensis, Isolates, Miltefosine, Susceptibility, Uptake, RNAseqhttp://www.sciencedirect.com/science/article/pii/S2211320718301684 |
| spellingShingle | Caroline R. Espada Rubens M. Magalhães Mario C. Cruz Paulo R. Machado Albert Schriefer Edgar M. Carvalho Valentín Hornillos João M. Alves Angela K. Cruz Adriano C. Coelho Silvia R.B. Uliana Investigation of the pathways related to intrinsic miltefosine tolerance in Leishmania (Viannia) braziliensis clinical isolates reveals differences in drug uptake International Journal for Parasitology: Drugs and Drug Resistance |
| title | Investigation of the pathways related to intrinsic miltefosine tolerance in Leishmania (Viannia) braziliensis clinical isolates reveals differences in drug uptake |
| title_full | Investigation of the pathways related to intrinsic miltefosine tolerance in Leishmania (Viannia) braziliensis clinical isolates reveals differences in drug uptake |
| title_fullStr | Investigation of the pathways related to intrinsic miltefosine tolerance in Leishmania (Viannia) braziliensis clinical isolates reveals differences in drug uptake |
| title_full_unstemmed | Investigation of the pathways related to intrinsic miltefosine tolerance in Leishmania (Viannia) braziliensis clinical isolates reveals differences in drug uptake |
| title_short | Investigation of the pathways related to intrinsic miltefosine tolerance in Leishmania (Viannia) braziliensis clinical isolates reveals differences in drug uptake |
| title_sort | investigation of the pathways related to intrinsic miltefosine tolerance in leishmania viannia braziliensis clinical isolates reveals differences in drug uptake |
| url | http://www.sciencedirect.com/science/article/pii/S2211320718301684 |
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